Structure-activity analysis of enolpyruvyl transferases

烯醇丙酮酰转移酶的结构-活性分析

• 批准号: 7393225
• 负责人: ERNST SCHONBRUNN
• 金额: $ 28.33万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7393225
• 关键词: Active Sites; American; Anabolism; Antibiotic Resistance; Antibiotics; Aromatic Amino Acids; Aromatic Compounds; Bacteria; Bacterial Infections; Binding; Biochemical Reaction; Catalysis; Cell Wall; Centers for Disease Control and Prevention (U.S.); Class; Commit; Communicable Diseases; Communities; Complex; Crystallography; Data; Depth; Development; Drug Delivery Systems; Drug Design; Enzyme Kinetics; Enzymes; Evaluation; Excitatory Postsynaptic Potentials; Family; Fluorescence Spectroscopy; Future; Goals; Growth; Kinetics; Knowledge; Laboratories; Ligands; Mammals; Methods; Molecular Cloning; Numbers; Organic Chemistry; Pathway interactions; Penicillins; Pharmaceutical Preparations; Plants; Property; Proteins; Proteomics; Qualifying; Range; Reaction; Research; Research Personnel; Role; Site; Solid; Structure; Structure-Activity Relationship; Techniques; Testing; Transferase; Work; analog; antimicrobial drug; base; design; functional group; inhibitor/antagonist; innovation; member; microbial; microorganism; microorganism growth; mutant; novel; pathogen; programs; shikimate; structural genomics

DESCRIPTION (provided by applicant): MurA and EPSPS are the only known members of the enolpyruvyl transferase family of enzymes. They are attractive targets for the development of new antibiotic drugs. The MurA reaction forms the first committed step in the biosynthesis of the bacterial cell wall. EPSP synthase is the sixth enzyme in the shikimate pathway towards the synthesis of aromatic amino acids and of other aromatic compounds in numerous microorganisms and plants. Both these pathways are absent from mammals but essential for microbial growth. The central hypothesis to the proposed project is that the use of selected mutant enzymes of MurA and EPSPS together with substrate, product and intermediate state analogs will allow for a complete understanding of the entire reaction pathway of enolpyruvyl transfer, starting from free enzyme to the enzyme products complex. The rationale for the proposed research is that once the prerequisites for the enzymatic reaction are identified, this information will be ultimately utilized to design potent inhibitors that selectively target both enzymes. In order to test our hypothesis and accomplish the objective of this project, two specific aims, which integrate techniques such as protein crystallography, enzyme kinetics, fluorescence spectroscopy, molecular cloning, and synthetic organic chemistry, will be pursued: (i) the identification of the prerequisites for the catalysis of enolpyruvyl transfer, and (ii) the identification of the prerequisites for the induced-fit mechanism. These studies will substantially contribute to our understanding of the relationship between the structure and activity of enolpyruvyl transferases. Moreover, they will provide a solid basis for the rational design of novel broad-spectrum antimicrobial drugs targeting these important enzymes.

描述（由申请人提供）：Mura和EPSP是酶的烯醇转移酶家族中唯一已知的成员。它们是开发新抗生素药物的有吸引力的靶标。 Mura反应在细菌细胞壁的生物合成中构成了第一个投入的步骤。 EPSP合酶是在许多微生物和植物中芳香氨基酸和其他芳族化合物合成的均匀途径中的第六个酶。 这两种途径都不是哺乳动物的，但对于微生物生长至关重要。拟议项目的中心假设是，使用Mura和EPSP的选定突变酶以及底物，产物和中间状态类似物将允许完全了解烯醇酸性L传递的整个反应途径，从自由酶开始，从自由酶开始到酶产物。拟议研究的基本原理是，一旦确定了酶促反应的先决条件，该信息将最终用于设计有效靶向这两种酶的有效抑制剂。 为了检验我们的假设并实现该项目的目标，两个具体的目的是整合诸如蛋白质晶体学，酶动力学，荧光光谱，分子克隆和合成有机化学等技术，将被追求：（i）用于鉴定的鉴定，以识别依次的鉴定。诱导拟合机制。 这些研究将大大有助于我们理解烯醇酸酯转移酶的结构和活性之间的关系。此外，它们将为针对这些重要酶的新型广谱抗菌药物的合理设计提供扎实的基础。